Previous studies have demonstrated low percentage of HL-60 cell differentiation with theophylline. The present study demonstrate that millimolar concentrations of the non-selective phosphodiesterase inhibitors theophylline, caffeine and isobutyl-methylxanthine all inhibit growth, induce substantial differentiation and elevation of both cAMP and cGMP in HL-60 cells. Selective inhibition of cAMP hydrolysis by Ro20-1724 was without effect. The guanylate cyclase stimulator sodium nitroprusside, which increased cGMP only poorly and also increased cAMP, produced growth inhibition but no differentiation. We put forward the hypothesis that elevation of both cAMP and cGMP above a critical level is necessary for significant cyclic nucleotide induced HL-60 cell differentiation.